Process of producing vaccine lymph



Patented Mar. 29, 19 38 UNITED STATES 2,112,507 PROCESS OF PRODUCING VACCINE LYMPH.

Klshiro Sukegawa, Koishikawa-ku, Tokyo, Japan No Drawing. Application June 29; 1935, Serial No. 29,175. In Japan December 28, 1933 Claims.

This invention relates to a process of producing vaccine lymph, and consists in adding to meat Juice a suitable decomposed albumin, e. g; pepton or amino-acid, and salt and sugar, neutralizing 5 it with colloidsol of sodium silicate or alkali-carbonate, adding hormone thereto, next adding ac tive elements oi. bird's ovum obtained during the process of it's hatching, thereby obtaining a cultural media, and then transmitting and cultivating the virus of smallpox in said cultural medium.

l The object of the invention is to cultivate the virus of smallpox by feeding them artificially, without relying upon the animal body as in the process heretofore practised, thereby obtaining,

in a simple and economical manner, the virus of smallpox necessary for the vaccination into the human body, whose reaction upon the skin is mild when inoculated upon the human body and whose protective power keeps long potent.

According to this invention, the liquid of meat juice is first added with a suitable decomposed albumin (e. g. pepton or amino-acid), salt and sugar such as grape-sugar or milk-sugar. The 5 mixture is neutralized by add ng thereto colloidsol of sodium silicate or alkali-carbonate. Then, to said neutralized liquid is added some sort of hormone body, e. g. such as pituitary body, or thyroid gland of hormone, or some chemical compound which has the equivalent efiect, and thereto is also added the efficacious element of bird's ovum obtained on the third day or on any day up to the twelfth day during the process of its hatching (e. g. in the case oi. a hens egg, embryonic 35 ectoblast tissue and plasma of the choroid urethral membrane, or an extract thereof), thereby obtaining a liquid or solid cultural medium.

Then, the virus of smallpox, namely the vaccine lymph separated from miscellaneous germs (the 0 vaccine lymph established according to the present regulation, or retrovaccination cow lymph established similarly) is transplanted and cultivated in the cultural media prepared in the manner mentioned above.

45 An example of the mode of carrying out this invention is as follows:-

To 100 parts of meat juice, there are added from 10 to 15 parts of the decomposed albumin, e. g. pepton or amino-acid, from 5 to '7 parts 0! 5 salt, and from 30 to 300 parts of grape sugar successively. Said liquid is then neutralized by adding from 0.1 to 0.2 part of colloidsol of sodium silicate. Then the same is admixed with the active element oi. suprarenal capsule, e. g. 5 parts 55 of 1000 fold solution of adrenalin, and a small quantity of the active elements of the ovum, i. e. embryonic ectoblast tissue, and plasma of the choroid urethral membrane obtained on the 8thday during the incubative process of a hens egg. Then the legal vaccine lymph is transplanted and is cultivated in the culture media thus prepared for 10 days in the incubator at 37 C. Then the product is filtered, and thereto is added 0.5% glycerinum of carbolic acid, thereby producing the pure, sterilized vaccine lymph. l0

In'carryingout my process, I may use aminoacid in place of pepton, milk-sugar in place of grape-sugar, egg of duck or quail in place of egg of hen, hormone of thyroid-gland or hypophysis in place of adrenalin, sodium silicate, with the 16 same satisfactory result. I may also use a solid culture media, which is made by adding Japanese gelatine to the above mentioned liquid nutriment.

A historical record of the methods of producing the vaccine lymph is as follows:- 2

(A) VARIOLATION This method was prevalent in India and China in ancient times and consists in inoculating the toxin oi smallpox directly upon the human body.

But this method often provokes actual case.

(B) VACCINATION This is known as Jenners method", for which the following four kinds of virus of smallpox are in use:

(1) Humanized vaccine (2) Original vaccine 40 This is a matter taken from the vesicle produced naturally on the cow's teat. But it is inconvenient inasmuch as this kind of vaccine is not always available.

(3) Revaccinated vaccine This is a matter taken from the vesicle produced by revaccinating humanized vaccine upon the body of a calf.

(4) Animal vaccine This is a matter taken from the vesicle produced by inoculating the original cow lymph upon the body of another calf.

Among the above-mentioned, (B), (2) and (4) are both improvements upon Jenner's method and it is the revaccinated in every country at present.

But all these cow'lymphs have severe reaction when they are inoculated upon the human body. 'In order to avoid such a defect, sometimes the rabbit is used as material.

And although in common use, vaccines which are produced upon the body of the calf are mingled with various kinds oi! germs and contain such animal organizations as the spore, lymph or scab of the calf, causing severe secondary reactions ,when inoculated upon the human body.

As for the miscellaneous bacteria mixed in the virus of smallpox, it has been proposedheretofore to sterilize them, mixing 1 part of vaccine lymph with 3 parts'of 80% water solution of glycerlne, and storing. it in a refrigerator for a period of one or two months. It has been also to add from 0.6 to 0.8% of carbolic acid to the glycerinated virus for the, same purp se. But I have found that all the above-mentioned existing methods were unable to make the virus perfectly nonbacilli. V Contrary to any of the old processes which rely upon vital body such as animal body in cultivating the virus, according to this invention the virus of smallpox is transplanted and cultivated upon I the artificial nutriment media, so that it is possible to easily and economically produce. the virus pure from any othergerms, which can make the human body immune with very slight reaction on the skin after theinoculation.

The vaccine, prepared according to the present invention, has been successfully subjected to a large number of experimental tests, using both animal and human subjects, and thus verifying the development of the virus of smallpox in the culture media of the invention, having been obtained in every case. In making these tests, hypodermic inoculation was employed. However, the vaccine of the present invention may also be used with the cutting method or by injection. The utility of the from the foregoing.

What I claim is:- v

1. A process for the production of vaccine lymph comprising adding an aminoacld-containing substance, salt and sugar to meat juice, neutralizing the product with a member of the group consisting of the colloidsols of sodium .silicate and sodium carbonate-adding adrenalin thereinvention is evident vaccine that isused a positive result then adding the active elements of a bird's I of embryonic ectoblast tissue and plasma of the urethral membrane obtained during the process of its hatchin thereby obtaining a cultural medium, and then transplanting and cultivating the virus of smallpox in said cultural medium.

2. A process for the production of vaccine lymph comprising addin peptone, salt and sugar to meat iuice, neutralizing the product with colloidsol of sodium silicate, adding adrenalin thereto, then addingthe active elements of a bird's ovum consisting ovum consisting of embryonic ectoblast tissue and plasma of the urethral membrane obtained during the process of its hatching, thereby obtaining a cultural ing and cultivating cultural medium.

the virus of smallpox in said 3. A process'for the production. or vaccine lymph comprising adding peptone, salt and sugar to meat juice, neutralizing the product with 'colloidsol of alkali carbonate, adding adrenalin thereto, then adding the active-elements of a bird's ovum consisting of embryonic ectobla'st tissue and plasma of the urethral membrane obmedium, and then transplanttained during the process of its hatching, thereby obtaining a cultural medium, and then transplanting and cultivating the virus of small pox in said cultural medium.

4. A process for the production of vaccine lymph comprising adding aminoacid, salt and sugar to meat juice, neutralizing the product with colloidsol of sodium thereto, then ad ng bird's ovum co isting of embryonic ectoblast tissue and plasma of the urethral membrane obtained during the process of its hatching, thereby obtaining a cultural medium, and then transplanting and cultivating the virus of smallpox in said cultural medium.

silicate, adding adrenalin 5. A process for the production of vaccine lymph comprising adding aminoacid, salt and sugar to meat juice, neutralizing the product with colloidsol of alkali carbonate, adding adrenalin thereto, then adding the active elements of a bird's cvum'consisting of embryonic ectoblast tissue and plasma of the urethral membrane obtained during the process of its hatching, thereby obtaining a cultural medium, and then transplanting and cultivating the virus of smallpox in said cultural medium.-

1 KISHIRO SUKEGAWA.

the active elements of a 

